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Biomedical subjects

S Ebisu

Publications and source records attributed to S Ebisu.

At least 109 records · Page 6Linked to original sources

Carbohydrate binding studies on the Bandeiraea simplicifolia I isolectins. Lectins which are mono-, di-, tri-, and tetravalent for N-acetyl-D-galactosamine.

Association constants for the binding of methyl alpha-D-galactopyranoside (methyl alpha-D-Galp) and methyl 2-acetamido-2-deoxy-alpha-D-galactopyranoside (methyl alpha-D-GalNAcp) to three Bandeiraea simplicifolia isolectins (A4, A2B2, B4) were determined by equilibrium dialysis and fluorescence enhancement measurements. The a and B subunits appear to have approximately the same Kassoc for methyl alpha-D-Galp: 1.45 X 10(4), 1.98 X 10(4), and 2.06 X 10(4) M-1 for A4, A2B2, and B4, respectively, as determined by equilibrium dialysis. Fluorescence enhancement measurements on B4 gave an association constant of 2.07 X 10(4) M-1 for methyl alpha-D-Galp and 1.87 X 10(3) M-1 for methyl beta-D-galp. By equilibrium dialysis, we were able to detect 3.3 (theory, 4.0) methyl alpha-D-GalNAcp binding sites for A4 (Kassoc = 1.87 X 10(5) M-1), 1.9 for A2B2 (Kassoc = 1.19 X 10(5) M-1), and were unable to detect any methyl alpha-D-GalNAcp binding sites for B4. However, four very weak methyl alpha-D-GalNAc binding sites for B4 were detected by fluorescence enhancement measurement (Kassoc = 1.26 X 10(2) M-1). Thus, the A subunit has an affinity for methyl alpha-D-GalNAc 3 orders of magnitude greater than the B subunit. Precipitation and hapten inhibition data are in accord with these binding measurements. Toward guaran and type B blood group substance, all isolectins precipitated the same amount of biopolymer. However, AB3, A2B2, and A3B, which are mono-, di-, and trivalent for alpha-D-GalNAcp, were differentially precipitated by type A blood group substance which contains alpha-D-GalNAcp-end groups. A3B precipitated the most, A2B2 less, and AB3 no type A substance. These isolectins should prove useful in studies evaluating the effect of valence on lectin-cell interaction.

ABO Blood-Group System↗

Eikenella corrodens adherence to human buccal epithelial cells.

The mechanism of Eikenella corrodens adherence to human buccal epithelial cells in vitro was studied. Initial experiments to determine the optimal conditions for adherence of E. corrodens to buccal epithelial cells showed that adherence was dependent on time, temperature, bacterial concentration, and pH. Different strains of E. corrodens varied in their ability to adhere, and strain 1073 showed the greatest ability in adherence. Strain 1073 was selected for studies of adherence mechanisms. Trypsin treatment or heating (100 degrees C, 10 min) of the bacterial cells abolished their capacity to adhere to buccal epithelial cells. Treatment of buccal epithelial cells with trypsin also abolished adherence of E. corrodens 1073, whereas neuraminidase treatment of buccal epithelial cells enhanced the adherence. The adherence was inhibited by ethylenediaminetetraacetic acid and restored by adding Ca2+. The adherence was remarkably inhibited by sugars containing D-galactose and n-acetyl-D-galactosamine. Treatment of neuraminidase-treated epithelial cells with sodium metaperiodate or alpha- and beta-galactosidase did not decrease the adherence. These data suggest that adherence of E. corrodens 1073 to human buccal epithelial cells may require the interaction of lectin-like proteins on the bacterial surface with galactose-like receptors on the surface of epithelial cells.

Bacteroides↗

A study of the mineral phase of pulp calcification.

Physico-chemical properties of pulp calcification were studied by means of X-ray microbeam diffraction, electron spin resonance (ESR), X-ray energy-dispersive analysis, and chemical analysis. The material was obtained from the second molar of the right mandible of a 25-year-old woman. X-ray diffraction and ESR analyses showed that calcium salts of the calcification are deposited in the form of apatitie, possibly carbonate-containing apatite. Furthermore, the mineral phase of the calcification was found to be similar to that of bone rather than that of dentin with regard to crystallinity and inorganic content. An unexpected finding was that high concentrations of iron were detected in some areas near the surface of the calcification. The question remains open as to what role, if any, iron may play in the formation of pulp calcification.

Adult↗

Mechanism of glucan-induced agglutination in Streptococcus mutans. I. Binding of radioactive glucan to whole cells of S. mutans OMZ-176.

The binding of radioactive glucan to Streptococcus mutans cells, which are agglutinated by dextrans, was examined. The glucan was synthesized from sucrose by extracellular glucosyltransferases from S. mutans FA-1 and was highly branched at C-3 and C-6 of D-glucose residues, containing 17% of a (1 leads to 3)inter-chain residues. Binding of glucan to whole cells of S. mutans OMZ-176, which were agglutinated by addition of glucan or Dextran T2000, was irreversible and followed saturation type kinetics; saturation was achieved at approximately 110 ng of glucan per ml. About 14 ng of glucan were bound per mg of the cells at the saturated concentration. The heated cells of this organism, however, had a relatively low ability of glucan-binding, compared with the freshly prepared and lyophilized cells. Binding to the heated cells was entirely of a non-saturation type. Binding of Dextran T2000 or T10 was determined by competition between the labeled glucan and unlabeled Dextrans for the binding site(s). Both Dextrans and glucan from S. mutans FA-1 were bound to the same site(s). Other organisms, which did not undergo glucan- and Dextran-induced agglutination, had a relatively lower ability of glucan-binding than S. mutans, which was agglutinated.

Agglutination↗

Activation of the alternative complement pathway by water-insoluble glucans of streptococcus mutans: the relation between their chemical structures and activating potencies.

Water insoluble, sticky glucan was synthesized by reacting glucosyltransferase obtained from the sulture filtrate of cariogenic Streptococcus mutans OMZ 176 on sucrose. This glucan (OMZ 176 glucan), that consisted of the backbone chains of consecutive alpha-1, 3 glucosidic linkages (65%) and the side chain of alpha-1, 6 glucosidic bonds (35%), showed an ability to activate the alternative pathway of the complment system in human serum. Water soluble glucans synthesized from the reaction of glycosyltransferases of S. mutans strain AHT, FA-I, and Ingbritt with sucrose, that have a high content of alpha-1,6 and a low content of alpha-1, 3 glucosidic linkages, were less effective to activate the complement system. Sephadex (G-25) and Dextran (T-2,000 and T-40), that consisted exclusively of alpha-1, 6 glucosidic linkages, did not significantly activate the complement system. Controlled Smith degradation products of OMZ 176 glucan, composed of the alpha-1, 3 linked glucose backbone chain alone, lost its activity. From these results it was concluded that both backbone chain of alpha-1, 3 glucose units and branched alpha-1, 6 glucose units were essential for the OMZ 176 glucan to activate the alternative pathway of the human complement system.

Aspergillus niger↗

Structural differences in fructans elaborated by streptococcus mutans and Strep. salivarius.

D-Fructans synthesized from sucrose by cell-free systems of strains of Streptococcus mutans and Strep. salivarius have been shown by methylation and enzymatic studies to have different glycosidic linkages. The cold water-insoluble D-fructans from Strep. mutans strain BHT and JC-1 have inulin-type structures consisting beta-(2 leads to 1)-D-fructofuranosidic linkages, with average repeating units of 8 and 27 sugar residues, respectively, whereas the water-soluble fructan from Strep. salivarius strain HHT has a levan-type branched structure consisting beta-(2 leads to 6)-D-fructofuranosidic linkages with an average repeating unit of 9 sugar residues. The solubility properties of these fructants are discussed on the basis of the structural differences.

Fructans↗

Isolation and purification of Flavobacterium alpha-1,3-glucanase-hydrolyzing, insoluble, sticky glucan of Streptococcus mutans.

Studies were made on the physical and chemical properties of polysaccharides synthesized by cell-free extracts of Streptococcus mutans, Streptococcus sanguis, and Streptococcus sp. and their susceptibilities to dextranases. Among the polysaccharides examined, insoluble glucans were rather resistant to available dextranase preparations, and the insoluble, sticky glucan produced by S. mutans OMZ 176, which could be important in formation of dental plaques, was the most resistant. By enrichment culture of soil specimens, using OMZ 176 glucans as the sole carbon source, an organism was isolated that produced colonies surrounded by a clear lytic zone on opaque agar plates containing the OMZ 176 glucan. The organism was identified as a strain of Flavobacterium and named the Ek-14 bacterium. EK-14 bacterium was grown in Trypticase soy broth, and an enzyme capable of hydrolyzing the OMZ 176 glucan was concentrated from the culture supernatant and purified by negative adsorption on a diethylaminoethyl-cellulose (DE-32) column and gradient elution chromatography with a carboxymethyl-cellulose (CM-32) column. The enzyme was a basic protein with an isoelectric point of pH 8.5 and molecular weight of 65,000. Its optimum pH was 6.3 and its optimal temperature was 42 C. The purified enzyme released 11% of the total glucose residues of the OMZ 176 glucan as reducing sugars and solubilized about half of the substrate glucan. The products were found to be isomaltose, nigerose, and nigerotriose, with some oligosaccharides. The purified enzyme split the alpha-1,3-glucan endolytically and was inactive toward glucans containing alpha-1,6, alpha-1,4, beta-1,3, beta-1,4, and/or beta-1,6 bonds as the main linkages.

Cell-Free System↗

Morphological characterization of tube-like structures in hypersensitive human radicular dentine.

OBJECTIVES: The objective of this work was to determine if there were differences in the distribution of tube-like structures within the tubules of biopsies from hypersensitive versus non-sensitive human cervical radicular dentine. METHOD: Ten pairs of dentine were made from the exposed sensitive and non-sensitive radicular dentine of nine adult patients complaining of dentine sensitivity. The biopsies were made with a small hollow diamond bur. The specimens were submitted to HCl-collagenase treatment to partially digest the surface to permit visualization of subsurface structures. Half of the specimens were embedded in resin to facilitate longitudinal sectioning, and the other half were processed with embedding for SEM and EDS observation. RESULTS: Biopsies from hypersensitive regions exhibited hollow, tube-like structures within the lumina of dentinal tubules in 75.8 +/- 9.4%) ( mean +/- S.D.) of the tubules. In biopsies of non-sensitive areas of the same teeth, these tube-like structures were only seen in 20.4 +/- 8.5% of the tubules. In specimens that were embedded prior to treatment with HCl-collagenase, resin tags penetrated many of the tubules from hypersensitive dentine but few of the tubules from non-sensitive dentine. CONCLUSION: It is speculated that tube-like structures found in hypersensitive dentine play a role in maintaining the patency of the tubules which may prevent physicochemical processes from spontaneously occluding the tubules, thereby maintaining dentine sensitivity.

Adult↗